Simultaneous enzyme overlay membrane (EOM)-based in situ zymography and immunofluorescence technique reveals cathepsin B-like activity in a subset of tumour vessels

2000 ◽  
Vol 114 (1) ◽  
pp. 63-68 ◽  
Author(s):  
Alexander Berndt ◽  
Jördis Johannesson ◽  
Michael Haas ◽  
Christoph Arkona ◽  
Detlef Katenkamp ◽  
...  
Keyword(s):  
Cathepsin B ◽  
Immunofluorescence Technique ◽  
In Situ Zymography

In Vivo Localization of Gelatinases (MMP-2 and -9) by in Situ Zymography with a Selective Gelatinase Inhibitor

2001 ◽  
Vol 287 (3) ◽  
pp. 766-774 ◽  
Author(s):  
Emma Pirilä ◽  
Päivi Maisi ◽  
Tuula Salo ◽  
Erkki Koivunen ◽  
Timo Sorsa
Keyword(s):  
In Situ Zymography

In Situ Proapoptotic Peptide-Generating Rapeseed Protein-Based Nanocomplexes Synergize Chemotherapy for Cathepsin-B Overexpressing Breast Cancer

2018 ◽  
Vol 10 (48) ◽  
pp. 41056-41069 ◽  
Author(s):  
Zhigao Wang ◽  
Rui Xue Zhang ◽  
Tian Zhang ◽  
Chunsheng He ◽  
Rong He ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cathepsin B ◽  
Rapeseed Protein

scholarly journals Cathepsin B-Overexpressed Tumor Cell Activatable Albumin-Binding Doxorubicin Prodrug for Cancer-Targeted Therapy

Pharmaceutics ◽  
2021 ◽  
Vol 14 (1) ◽  
pp. 83
Author(s):  
Hanhee Cho ◽  
Man Kyu Shim ◽  
Suah Yang ◽  
Sukyung Song ◽  
Yujeong Moon ◽  
...  
Keyword(s):  
Tumor Cell ◽  
Cathepsin B ◽  
Half Life ◽  
Albumin Binding ◽  
Normal Tissues ◽  
Cleavable Peptide ◽  
Tumor Accumulation ◽  
Maleimide Group ◽  
Cancer Targeted Therapy

Prodrugs are bioreversible medications that should undergo an enzymatic or chemical transformation in the tumor microenvironment to release active drugs, which improve cancer selectivity to reduce toxicities of anticancer drugs. However, such approaches have been challenged by poor therapeutic efficacy attributed to a short half-life and low tumor targeting. Herein, we propose cathepsin B-overexpressed tumor cell activatable albumin-binding doxorubicin prodrug, Al-ProD, that consists of a albumin-binding maleimide group, cathepsin B-cleavable peptide (FRRG), and doxorubicin. The Al-ProD binds to in situ albumin, and albumin-bound Al-ProD indicates high tumor accumulation with prolonged half-life, and selctively releases doxorubicin in cathepsin B-overexpressed tumor cells, inducing a potent antitumor efficacy. Concurrently, toxicity of Al-ProD toward normal tissues with innately low cathepsin B expression is significantly reduced by maintaining an inactive state, thereby increasing the safety of chemotherapy. This study offers a promising approach for effective and safe chemotherapy, which may open new avenues for drug design and translational medicine.

Evaluation of matrix metalloproteinase-2 (mmp-2) activity with film in situ zymography for improved cytological diagnosis of Breast tumors

Breast Cancer ◽  
2006 ◽  
Vol 13 (3) ◽  
pp. 272-278 ◽  
Author(s):  
Akira Fujiwara ◽  
Eri Shibata ◽  
Hitomi Terashima ◽  
Akemi Shishido ◽  
Junko Nishiki ◽  
...  
Keyword(s):  
Matrix Metalloproteinase ◽  
Breast Tumors ◽  
Matrix Metalloproteinase 2 ◽  
Cytological Diagnosis ◽  
Film In Situ Zymography ◽  
In Situ Zymography

Abstract 212: Fractalkine Neutralization Improves Cardiac Function After Myocardial Infarction

Hypertension ◽  
2014 ◽  
Vol 64 (suppl_1) ◽  
Author(s):  
Xiaosong Gu ◽  
Jiang Xu ◽  
Xiao-Ping Yang ◽  
Edward Peterson ◽  
Pamela Harding
Keyword(s):  
Myocardial Infarction ◽  
Cardiac Function ◽  
Macrophage Infiltration ◽  
Receptor Subtype ◽  
Cross Sectional ◽  
Gelatinase Activity ◽  
Ep4 Receptor ◽  
In Situ Zymography ◽  
Circulating Levels

Background: Circulating levels of the chemokine fractalkine (FKN) are increased in patients with chronic heart failure (HF) and our studies show that aged mice lacking the prostaglandin E2 EP4 receptor subtype (EP4 KO) have increased cardiac FKN coupled with a phenotype of dilated cardiomyopathy. However, whether FKN is a causal factor for HF is not well established. Hypothesis: FKN contributes to the pathogenesis of HF post myocardial infarction (MI) and EP4 KO mice have a better response to anti-FKN treatment due to elevated FKN levels. Methods: Male EP4 KO mice and wild type (WT) littermates underwent surgery to induce MI. Mice were treated with an anti-FKN antibody (40μg/kg/day, ip) or IgG immediately after MI and echocardiography was performed 2 wks post MI. Hearts were excised for infarct size determination, myocyte cross-sectional area (MCSA) and interstitial collagen fraction (ICF) determined by morphometric analysis and macrophage infiltration using immunohistochemistry. Results: Anti-FKN treatment improved cardiac function and prevented remodeling (Table). In situ zymography revealed that gelatinase activity was markedly reduced by anti-FKN treatment in both strains. Moreover, anti-FKN treatment tended to improve survival in EP4 KO mice (p = 0.06, n=20). Conclusions: (1) FKN contributes to the pathogenesis of HF and anti-FKN treatment improves cardiac function and reduces cardiac remodeling. This may be related to reduced macrophage infiltration and decreased gelatinase activity.(2) Contrary to our hypothesis, EP4 KO mice do not have an enhanced response to anti-FKN treatment.

scholarly journals MMP Activity in the Hybrid Layer Detected with in situ Zymography

2012 ◽  
Vol 91 (5) ◽  
pp. 467-472 ◽  
Author(s):  
A. Mazzoni ◽  
F.D. Nascimento ◽  
M. Carrilho ◽  
I. Tersariol ◽  
V. Papa ◽  
...  
Keyword(s):  
Hybrid Layer ◽  
In Situ Zymography

scholarly journals Influence of Therapeutic Hypothermia on Matrix Metalloproteinase Activity after Traumatic Brain Injury in Rats

2005 ◽  
Vol 25 (11) ◽  
pp. 1505-1516 ◽  
Author(s):  
Jessie S Truettner ◽  
Ofelia F Alonso ◽  
W Dalton Dietrich
Keyword(s):  
Brain Injury ◽  
Therapeutic Hypothermia ◽  
Brain Injuries ◽  
Brain Temperature ◽  
Sprague Dawley ◽  
Beneficial Effects ◽  
Sprague Dawley Rats ◽  
In Situ Zymography ◽  
Metalloproteinase Activity

Recent evidence suggests that matrix metalloproteinases (MMPs) contribute to acute edema and lesion formation following ischemic and traumatic brain injuries (TBI). Experimental and clinical studies have also reported the beneficial effects of posttraumatic hypothermia on histopathological and behavioral outcome. The purpose of this study was to determine whether therapeutic hypothermia would affect the activity of MMPs after TBI. Male Sprague-Dawley rats were traumatized by moderate parasagittal fluid-percussion (F-P) brain injury. Seven groups ( n = 5/group) of animals were investigated: sham-operated, TBI with normothermia (37°C), and TBI with hypothermia (33°C). Normothermia animals were killed at 4, 24, 72 h and 5 days, and hypothermia animals at 24 or 72 h. Brain temperature was reduced to target temperature 30 mins after trauma and maintained for 4 h. Ipsilateral and contralateral cortical, hippocampal, and thalamic regions were analyzed by gelatin and in situ zymography. In traumatized normothermic animals, TBI significantly ( P<0.005) increased MMP-9 levels in ipsilateral (right) cortical and hippocampal regions, compared with contralateral or sham animals, beginning at 4 h and persisting to 5 days. At 1, 3, and 5 days after TBI, significant increases in MMP-2 levels were observed. In contrast to these findings observed with normothermia, posttraumatic hypothermia significantly reduced MMP-9 levels. Hypothermic treatment, however, did not affect the delayed activation of MMP-2. Clarifying the mechanisms underlying the beneficial effects of posttraumatic hypothermia is an active area of research. Posttraumatic hypothermia may attenuate the deleterious consequences of brain trauma by reducing MMP activation acutely.

High resolution in situ zymography reveals matrix metalloproteinase activity at glutamatergic synapses

Neuroscience ◽  
2009 ◽  
Vol 158 (1) ◽  
pp. 167-176 ◽  
Author(s):  
M. Gawlak ◽  
T. Górkiewicz ◽  
A. Gorlewicz ◽  
F.A. Konopacki ◽  
L. Kaczmarek ◽  
...  
Keyword(s):  
High Resolution ◽  
Matrix Metalloproteinase ◽  
Glutamatergic Synapses ◽  
In Situ Zymography ◽  
Metalloproteinase Activity

In Situ Zymography

2010 ◽  
pp. 271-277 ◽  
Author(s):  
Sarah J. George ◽  
Jason L. Johnson
Keyword(s):  
In Situ Zymography

scholarly journals Langerhans Cells Express IL-17A in the Epidermis of Chromoblastomycosis Lesions

2017 ◽  
Vol 2 (2) ◽  
pp. 1-8 ◽  
Author(s):  
Aline Alves de Lima Silva ◽  
Paulo Ricardo Criado ◽  
Ricardo Spina Nunes ◽  
Luciane Kanashiro-Galo ◽  
Maria Irma Seixas Duarte ◽  
...  
Keyword(s):  
Langerhans Cells ◽  
Subcutaneous Tissue ◽  
Innate Immune ◽  
Lesion Group ◽  
Control Group ◽  
Histopathological Diagnosis ◽  
Immunofluorescence Technique ◽  
Double Immunostaining ◽  
First Time

Chromoblastomycosis (CBM) is a chronic fungal infection that affects skin and subcutaneous tissue, and little is known about the immunological aspects of such lesions. We have previously described the high expression of IL-17 in this group. Understanding the innate immune response of patients with CBM would improve the knowledge of its immunopathogenesis and contribute to the most appropriate therapies. Nineteen biopsies of verrucous form were obtained from patients with clinical and histopathological diagnosis of CBM, without treatment. This was done with a double immunostaining with conventional immunohistochemistry and immunofluorescence technique as well as confocal microscopy to detect Langerin and IL-17 expression. All of the specimens that were analyzed showed expression of Langerin in the epidermis - the same as the control group. However, only the CBM group presented cells expressing CD207 in the dermis. Interestingly, the coexpression of IL-17 and Langerin was visualized along the epidermis and dermis in 100% of the lesion group. We demonstrated for the first time in situ coexpression of IL-17 and Langerin (CD207) in epidermal cells of patients with CBM and speculated on their role as IL-17-producing cells or whether they could be a new subpopulation of dendritic cells distinct from Langerhans cells.

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